自体外周血CD34^＋造血干细胞移植治疗多发性骨髓瘤

[目的]探讨自体外周血CD+34造血干细胞移植治疗多发性骨髓瘤的可行性。[方法]应用CliniMACS系统分选纯化 患者外周血CD+34造血干细胞,用流式细胞仪检测分选纯化 结果。选马法兰200mg/m2作为预处理方案。[结果]分选纯化结 果,单个核细胞数为3.7136×108,活性率96.6％,CD+34细胞数 3.68018×108,即5.52×106/kg。移植后患者骨髓像获得完全缓 解。移植相关并发症少且易于控制。[结论]自体外周血CD+34 造血干细胞移植治疗多发性骨髓瘤是安全、有效、可行的 。     

纯化的自体CD34~+细胞移植治疗恶性血液病临床对照观察

 目的　分析CD34+ 纯化自体造血干细胞移植治疗恶性血液病的临床疗效。方法　 11例病人进行了CD34+ 纯化自体造血干细胞移植 ,8例患者进行了未分选的自体造血干细胞移植 ,动员方案为CTX+Vp 16联合G CSF。CD34+ 细胞分选采用CliniMACS系统。结果　分选后CD34+ 细胞纯度达到(88.99± 6 .88) % ,回收率达到 (6 9.0 6± 17.0 7) %。 11例分选患者感染率为 (5 / 11例 ) ,复发率为 (1/ 11例 ) ,死亡率为 (3/ 11例 )。外周血中性粒细胞 >0 .5× 10 9/L时间为 (11± 1)天 ,血小板 >2 0× 10 9/L为(13± 3)天 ,8例未行CD34+ 分选患者的感染率 (4/ 8例 ) ,复发率为 (3/ 8例 ) ,死亡率为 (5 / 8例 )。外周血中性粒细胞 >0 .5× 10 9/L时间 (11± 1)天和血小板 >2 0× 10 9/L(14± 4 )天。结论　利用CliniMACs系统体外进行外周血CD34+ 细胞的富集与纯化纯度、回收率均满意 ,自体移植后造血功能重建顺利 ;与对照相比造血重建时间、感染率...     

自体外周血CD_(34)~ 造血干细胞移植治疗多发性骨髓瘤

[目的]探讨自体外周血CD 造血干细胞移植治疗多发性骨髓瘤的可行性。[方法]应用CliniMACS系统分选纯化患者外周血CD 造血干细胞 ,用流式细胞仪检测分选纯化结果。选马法兰200mg/m2 作为预处理方案。[结果]分选纯化结果 ,单个核细胞数为3.7136×108,活性率96.6% ,CD 细胞数3.68018×108,即5.52×106/kg。移植后患者骨髓像获得完全缓解。移植相关并发症少且易于控制。[结论]自体外周血CD 造血干细胞移植治疗多发性骨髓瘤是安全、有效、可行的。     

外周血CD_(34)~+细胞检测对外周血干细胞采集结果及时机选择的意义

 目的　研究外周血CD+34 细胞数对采集结果的意义,并探索可用于临床指导外周干细胞采集时机选择的参考阈值。方法　2007年1月至2009年12月共57例次自体造血干细胞移植动员采集患者,以环磷酰胺（CTX）化疗+粒细胞集落刺激因子（G-CSF）（5～10 μg／kg）动员,COBE分离仪（Spectra Version 6）行外周血造血干细胞采集,应用流式细胞术监测外周血中CD+34 细胞绝对计数。结果　采集产品单个核细胞（MNC）中位数4.6×108／kg（0.3×108／kg～10.5×108／kg）,CD+34 细胞中位数2.4×106／kg（0.16×106／kg～34.9×106／kg）,外周血CD+34 细胞数是产品MNC和CD+34 细胞总量唯一相关指标,外周血白细胞（WBC）与采集产品MNC和CD+34 细胞数无关。进一步分析提示外周血CD+34 计数≥15／μl,单次采集效率提高,CD+34 细胞采集量达1×106／kg和2×106／kg比例为81 %和60 %,采集产品MNC和CD+34 总数明显提高。提示外周血CD+34 细胞数15／μl可作为启动采集。ROC分析发现外周血CD+34 细胞 25（26.5～28.6）／μl,单次采集足量CD+34 细胞概率最大。结论　外周血CD+34 细胞计数是外周血自体干细胞采集重要的相关指标,CD+34 细胞 15／μl可作为采集时机选择的阈值。     

正常供者外周血造血干细胞动员效果及影响因素

 目的　探讨粒细胞集落刺激因子（G-CSF）动员正常供者外周血造血干细胞的效果、毒副作用及影响因素。方法　对59名异基因造血干细胞正常供者采用G-CSF 皮下注射3 ~ 5 d,使用COBE Spectra血细胞分离机采集外周血干细胞,流式细胞术检测采集物中CD+34细胞数。结果　所有供者第一次采集的单个核细胞（MNC）及CD+34细胞量平均值分别为4.4（1.12～13.06）×108／kg供者体重及3.78（1.14～12.92）×106／kg供者体重。患者不良反应轻微。男性供者、年龄小于45岁者及采集前白细胞计数高者采集所得CD+34细胞数较高。结论　G-CSF作为正常供者动员剂安全有效。患者性别、年龄及采集前白细胞计数可作为预测因素。     

自体CD34~+细胞移植治疗晚期肿瘤

目的　采用CD34+ 细胞体外分选技术对晚期肿瘤患者进行自体CD34+ 细胞移植 ,以降低自体移植后肿瘤复发率。方法　对 15例Ⅲ～Ⅵ期肿瘤患者 (多发性骨髓瘤 11例 ,乳癌 2例 ,非霍奇金淋巴瘤和髓母细胞瘤各 1例 )采用CliniMACS临床型细胞富集仪 ,利用磁性分选技术收集CD34+和CD34-细胞组分 ,患者于预处理后 ,输注分选后的CD34+ 细胞。结果　CD34+ 细胞体外纯化富集可使CD34-细胞获得 2 .0～ 5 .0个对数的去除 ;回输CD34+ 细胞中位数为 2 .4× 10 6/kg,CD34+ 细胞回收率为 6 4 % ,纯度为 98.2 % ;移植后白细胞恢复至 >1.0× 10 9/L和血小板 >2 0× 10 9/L的天数 (中位数 )分别为 14d和 13d。患者总体生存率 6 6 .7% (10 / 15 ) ,无疾病生存率 5 3.3% (8/ 15 )。结论　CD34+ 细胞移植后获得迅速、稳定的造血重建。体外CD34+ 细胞纯化富集后移植可望提高晚期肿瘤患者自体移植疗效。     

中剂量环磷酰胺为主的联合化疗加同一剂量G-CSF动员恶性血液病患者自体外周血干细胞

目的:观察中剂量环磷酰胺(CTX)为主的联合化疗加G蛳CSF对恶性血液病患者自体外周血造血干细胞(APBSC)的动员效果。方法:31例患者接受中剂量CTX 2.2 g/m2(1.8 g/m2~3.0 g/m2)联合VP16(600 mg ~ 800 mg)或Ara蛳C(1.0 g/m2 ~ 2.0 g/m2)化疗,WBC降至最低值后开始皮下注射G蛳CSF 300 μg/d直至采集结束。WBC≥(3.0~5.0)×109/L时开始采集,当单个核细胞(MNC)累计≥3.8×108/kg或CD+34细胞≥2.0×106/kg时停止采集。结果:采集次数为(2.9±1.0)次,G蛳CSF持续应用时间为(7.4±2.0)d,采集到的MNC细胞数为(5.53±2.54)×108/kg,CD+34细胞数为(9.46±7.24)×106/kg,CFU蛳GM(46.02±70.58)×104/kg。全部移植患者造血功能均获满意重建。结论:中剂量CTX为主的联合化疗加同一剂量G蛳CSF对血液病患者的APBSC动员是安全、有效的。     

紫杉醇联合重组人粒细胞集落刺激因子动员 乳腺癌患者外周血干细胞的效果及影响因素

 目的研究紫杉醇联合重组人粒细胞集落刺激因子（rhG-CSF）动员乳腺癌患者外周血干细胞（peripheral blood stem cell,PBSC）的效果及影响因素分析。方法2006年2月至2009年6月我科收治行紫杉醇动员的26例乳腺癌患者,紫杉醇（PTX,175 mg/m2 持续静脉滴注24 h）化疗后,白细胞降至1.0×109/L左右时使用rhG-CSF 5 μg /(kg·d) 动员至采集结束。并进一步分析患者年龄,化疗后白细胞最低数,采集前各类血细胞数,术后分期以及既往化疗等因素对采集单个核细胞（mononuclear cell,MNC）、CD34+细胞数的影响。结果白细胞计数于紫杉醇化疗后中位7d降至1.0×109/L 左右,皮下注射rhG-CSF中位4d进行外周造血干细胞采集,采集总MNC平均(7.89±1.45)×108/kg,采集总CD34+细胞平均(4.88±1.54)×106/kg。年龄与采集CD34+细胞数显著相关。而其他因素对MNC及CD34+细胞数均无显著影响（P>0.05）。所有患者均未出现严重不良反应。结论PTX（175 mg/m2 持续静脉滴注24h）联合rhG-CSF为转移性乳腺癌患者动员的有效安全方案。患者年龄显著影响CD34+细胞的采集数量。     

外周血造血干细胞的动员、采集和移植后造血重建

目的对13例健康供者及患者外周血造血干细胞的动员和采集效果进行分析。方法健康供者用rhG-CSF动员;患者采用强化疗加rhG-CSF或和rhGM-CSF动员,COBESpectro血细胞分离机全自动造血干细胞程序采集单个核细胞。结果13例移植中8例一次采集成功,1例(即首例)采集4次,余者采集2次。采集后的MNC6×108/kg～12×108/kg,CD3+4细胞3×106/kg～34×106/kg。移植后全部患者造血均获重建,异基因外周血造血干细胞移植一个月后DNA指纹图均提示植活。结论rhG-CSF可作为健康供者的安全有效动员剂,强化疗加rhG-CSF或/和rhGM-CSF是白血病、淋巴瘤自体外周血造血干细胞移植的有效动员方法之一。     

60例实体肿瘤患者外周血干细胞的动员和采集

目的:对60例实体肿瘤患者造血干细胞的动员和采集效果进行分析。方法:采用常规化疗联合造血因子动员剂方法动员肿瘤患者外周血干细胞,CS-3000plus血细胞分离仪和程控冷冻降温方法采集、冷冻保存干细胞。结果:60例患者平均年龄41岁(24～57),Ⅰ期3.4%,Ⅱ期25.0%,Ⅲ期38.3%,Ⅳ期33.3%;肿瘤负荷情况,CR46.6%,PR31.6%。平均采集次数2.6次,MNC5.6×108/kg,CFU-GM2.4×105/kg,BFU-E1.7×104/kg,CD34+10.1×106/kg,CFU-GM第1次收获率50.0%,第2次36.0%。各病种间除CD34+外均无统计学差异。恶性淋巴瘤组CD34+细胞数高于其他病种。一次动员成功率90.0%。干细胞冷冻复苏后平均MNC回收率77.0%,CFU-GM回收率69.0%,BFU-E回收率72.0%。各病种之间回收率无差异。结论:常规化疗联合造血干细胞因子是实体肿瘤外周血干细胞移植的一种有效动员剂。     

Biomarkers in clear cell renal cell carcinoma

The treatment of advanced renal cell carcinoma (RCC) has evolved significantly following the identification of the von Hippel–Lindau (VHL) gene and the function of its protein, and subsequent development of antiangiogenic therapies. A series of clinical trials resulted in the approval of three new agents with significant activity in this disease. Additional studies are now underway to identify subsets of patients most likely to benefit. This article reviews the current therapy for advanced RCC and the development of biomarkers in RCC. This requires the identification of disease characteristics at a clinical, genetic and molecular level associated with response and/or surrogate measures of clinical benefit. Currently, a variety of prognostic factors (lactate dehydrogenase, performance status, disease-free interval, hemoglobin and calcium levels) are utilized to predict the survival of RCC patients. The use of validated biomarkers in either serum/plasma, urine or tissue could enhance this process, as well as define at the molecular and genetic levels, factors associated with response to therapy and/or the development of resistance. Examples include plasma VEGF levels, VHL gene mutation status and carbonic anhydrase IX levels in tumor tissue, among others. Validation of such biomarkers is crucial in order for them to be clinically useful.     

Biomarkers in clear cell renal cell carcinoma

The treatment of advanced renal cell carcinoma (RCC) has evolved significantly following the identification of the von Hippel-Lindau (VHL) gene and the function of its protein, and subsequent development of antiangiogenic therapies. A series of clinical trials resulted in the approval of three new agents with significant activity in this disease. Additional studies are now underway to identify subsets of patients most likely to benefit. This article reviews the current therapy for advanced RCC and the development of biomarkers in RCC. This requires the identification of disease characteristics at a clinical, genetic and molecular level associated with response and/or surrogate measures of clinical benefit. Currently, a variety of prognostic factors (lactate dehydrogenase, performance status, disease-free interval, hemoglobin and calcium levels) are utilized to predict the survival of RCC patients. The use of validated biomarkers in either serum/plasma, urine or tissue could enhance this process, as well as define at the molecular and genetic levels, factors associated with response to therapy and/or the development of resistance. Examples include plasma VEGF levels, VHL gene mutation status and carbonic anhydrase IX levels in tumor tissue, among others. Validation of such biomarkers is crucial in order for them to be clinically useful.     

Non-small and small cell lung carcinoma cell lines exhibit cell type-specific sensitivity to edelfosine-induced cell death and different cell line-specific responses to edelfosine treatment

The unique signal transduction pathways that distinguish non-small cell lung carcinoma (NSCLC) from small cell lung carcinoma (SCLC) are poorly understood. We investigated the ability of edelfosine, an inhibitor of phosphatidylinositol-specific phospholipase C (PLC) to inhibit cell viability among four NSCLC cell lines and four SCLC cell lines. The differential sensitivity of cells to edelfosine's cytostatic and cytotoxic effects has been attributed to edelfosine-induced changes in the activities of many enzymes, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinases (ERK), p38 kinase, and poly(ADP-ribose) polymerase (PARP). To investigate the role of these enzymes in edelfosine-induced cytotoxicity, we correlated edelfosine-induced changes in enzyme activity and cell viability among the different NSCLC and SCLC cell lines. We found that NSCLC cells are much more susceptible to the cytotoxic effects of this drug than are SCLC cells. Three out of the four edelfosine-sensitive NSCLC cell lines (NCI-H157, NCI-H520, NCI-H522) exhibit G2/M arrest, significant apoptosis and some degree of JNK activation in response to drug treatment. In contrast, none of the SCLC cell lines exhibit edelfosine-induced G2/M arrest or significant apoptosis. A comparison of the edelfosine-induced effects among the sensitive and resistant lung cancer lines indicates that there is little correlation between edelfosine-induced cytotoxicity and altered activities of JNK, ERK, p38, or cleavage of PARP. These results demonstrate that edelfosine-induced changes in JNK, ERK, p38, or PARP are not good predictors of cell susceptibility to edelfosine-induced cytotoxicity. Thus, edelfosine-induced inactivation of PLC may disrupt signaling cascades downstream of PLC that are unique to individual cellular environments. These findings also identify edelfosine as one of the few potential chemotherapeutic agents that has a greater cytotoxic effect against NSCLC cells than SCLC cells.     

桥接整合因子1通过AKT-mTOR通路诱导非小细胞肺癌H1975细胞周期阻滞

目的：研究桥接整合因子1（bridging intergrator 1,Bin1）基因过表达后对非小细胞肺癌细胞株H1975细胞周期的影响及其作用机制。方法：构建携带Bin1基因的CMV-MCS-GFP-SV40-Neomycin-Bin1质粒,并转染H1975细胞（Bin1+组）,另设置空白质粒转染组（Bin1-组）及空白对照组（Ctrl组）,利用RT-PCR和Western blotting分别检测3组细胞中Bin1在mRNA和蛋白质水平的表达情况。流式细胞术检测不同处理组H1975细胞周期的变化,Western boltting分别检测各组中AKT、mTOR磷酸化水平及细胞周期相关蛋白（周期蛋白D1、CDK4、Rb）的表达情况。结果：与Bin1-组、Ctrl组比较,Bin1+组H1975细胞中Bin1在mRNA、蛋白水平表达明显上调（均P<0.05）; H1975细胞阻滞在G1期\[（60.53±1.89）% vs（46.14±1.56）%、（47.33±2.07）%,均P<0.05\]; Bin1+组H1975细胞内p-AKT、p-mTOR表达下调（均P<0.05）,AKT、mTOR表达变化无统计学差异（P>0.05）;周期蛋白D1、CDK4的表达量均明显下调(P<0.05),Rb表达量明显增加（P<0.05）。结论：Bin1基因在H1975细胞株过表达后明显诱导细胞周期阻滞,其机制可能是通过抑制AKT-mTOR通路及其细胞周期相关蛋白实现的。     

Inhibition of S-adenosylhomocysteine hydrolase decreases cell mobility and cell proliferation through cell cycle arrest

S-adenosylhomocysteine hydrolase (AHCY) hydrolyzes S-adenosylhomocysteine to adenosine and l-homocysteine, and it is already known that inhibition of AHCY decreased cell proliferation by G2/M arrest in MCF7 cells. However, the previous study has not indicated what mechanism the cell cycle arrest is induced by. In this study, we aimed to investigate the different cell cycle mechanisms in both p53 wild-typed MCF7 and p53 mutant-typed MCF7-ADR by suppressing AHCY. We extensively proved that AHCY knockdown has an anti-proliferative effect by using the WST-1 assay, BrdU assay, and cell cytometry analysis and an anti-invasive, migration effect by wound-healing assay and trans-well analysis. Our study showed that down-regulation of AHCY effectively suppressed cell proliferation by regulating the MEK/ERK signaling pathway and through cell cycle arrests. The cell cycle arrest occurred at the G2/M checkpoint by inhibiting degradation of cyclinB1 and phosphorylation of CDC2 in MCF7 cells and at the G1 phase by inhibiting cyclinD1 and CDK6 in MCF7-ADR cells. Finally, we determined that AHCY regulates the expression of ATM kinase that phosphorylates p53 and affects to arrest of G2/M phase in MCF7 cells. The findings of this study significantly suggest that AHCY is an important regulator of cell proliferation through different mechanism in between MCF7 and MCF7-ADR cells as p53 status.